Severevocalfold(VF)mucosalinjuryanditsassociatedvoiceimpairment(dysphonia)isadebilitatingand challengingclinicalproblemwithfewtreatmentoptions,noneofwhichareconsistentlyeffective.Inpatients withsuchtissueimpairmentorloss,anengineeredVFmucosawouldprovideabiomechanicallyappropriate tissuegraftforvoicerestoration.Duringourpreviousfundingcycle,weengineeredsuchaVFmucosavia organotypiccultureofprimaryvocalfoldfibroblastsandepithelialcellsfromhumandonors.Inaseriesof preclinicalexperiments,theengineeredmucosarecapitulatedthestructureandphysiologicfunctionofnative humantissue.Inthecurrentapplication,weproposeanaturalextensionofourpriorworkbyadvancing characterizationoftheinvitrotissueengineeringprocessatthebiologicalsystemlevel(Aim1),studying remodelingandtheadaptiveimmuneresponsetotheengineeredmucosafollowinginvivoheterotopic implantation(Aim2),andstudyinglong-termphysiologicandimmunologicoutcomesfollowinginvivo orthotopicimplantation(Aim3).Theseexperimentswillgenerateimportantpreclinicalsafetyandefficacydata. Wewillcombinetheplannedexperimentalworkwithregulatoryandclinicalmanufacturingmilestones(Aim4), withafinalgoalofholdingapre-InvestigationalNewDrugmeetingwiththeFDAbytheendofthegrant.The tissueengineeringtechnologyoutlinedinthisapplicationhasthepotentialtotransformtreatmentofthemost recalcitrantVFtissuedisordersencounteredintheclinic.